LOCUS       sequence-494445-        6179 bp    DNA     circular SYN 02-DEC-2025
DEFINITION  Retroviral expression vector for secreted E. coli BirA, C-terminal 
            FLAG tag.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6179)
  AUTHORS   Heiden R, Hannig L, Kuo CJ, Ergun S, Braunger BM, Vallon M
  TITLE     WNT7A/B assemble a GPR124-RECK-LRP5/6 co-receptor complex to 
            activate beta-catenin signaling in brain endothelial cells.
  JOURNAL   J Biol Chem. 2025 Sep 4:110682. doi: 10.1016/j.jbc.2025.110682.
  PUBMED    40914247
REFERENCE   2  (bases 1 to 6179)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 2, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "J Biol 
            Chem. 2025 Sep 4:110682. doi: 10.1016/j.jbc.2025.110682."
COMMENT     Sequence Label: pBABE-Puro-BirA
FEATURES             Location/Qualifiers
     source          1..6179
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             8..477
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     misc_feature    542..741
                     /label=MMLV Psi
                     /note="packaging signal of Moloney murine leukemia virus 
                     (MMLV)"
     CDS             942..1358
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
     primer_bind     1288..1310
                     /label=pLXSN 5'
                     /note="Murine stem cell virus, forward primer. Also called 
                     MSCV"
     primer_bind     1326..1342
                     /label=pBABE 5'
                     /note="Psi packaging signal, forward primer for pBABE 
                     vectors"
     regulatory      1393..1402
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1459..2418
                     /codon_start=1
                     /product="E. coli biotin protein ligase"
                     /label=BirA
                     /translation="KDNTVPLKLIALLANGEFHSGEQLGETLGMSRAAINKHIQTLRDW
                     GVDVFTVPGKGYSLPEPIQLLNAKQILGQLDGGSVAVLPVIDSTNQYLLDRIGELKSGD
                     ACIAEYQQAGRGRRGRKWFSPFGANLYLSMFWRLEQGPAAAIGLSLVIGIVMAEVLRKL
                     GADKVRVKWPNDLYLQDRKLAGILVELTGKTGDAAQIVIGAGINMAMRRVEESVVNQGW
                     ITLQEAGINLDRNTLAAMLIRELRAALELFEQEGLAPYLSRWEKLDNFINRPVKLIIGD
                     KEIFGISRGIDKQGALLLEQDGIIKPWMGGEISLRSAEK"
     CDS             2425..2448
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     primer_bind     complement(2509..2529)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2514..2843
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2694..2829
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2853..3452
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(2853..2872)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     3349..3369
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     LTR             3673..4142
                     /note="long terminal repeat from Moloney murine leukemia 
                     virus"
     primer_bind     complement(4272..4289)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4443..5031)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4523..4542)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5191..5982,5983..6051))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5814..5833
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6052..6156)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctgcagcctg aatatgggcc aaacaggata tctgtggtaa gcagttcctg ccccggctca
       61 gggccaagaa cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt
      121 tcctgccccg gctcagggcc aagaacagat ggtccccaga tgcggtccag ccctcagcag
      181 tttctagaga accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc
      241 ttatttgaac taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga
      301 gctcaataaa agagcccaca acccctcact cggggcgcca gtcctccgat tgactgagtc
      361 gcccgggtac ccgtgtatcc aataaaccct cttgcagttg catccgactt gtggtctcgc
      421 tgttccttgg gagggtctcc tctgagtgat tgactacccg tcagcggggg tctttcattt
      481 gggggctcgt ccgggatcgg gagacccctg cccagggacc accgacccac caccgggagg
      541 caagctggcc agcaacttat ctgtgtctgt ccgattgtct agtgtctatg actgatttta
      601 tgcgcctgcg tcggtactag ttagctaact agctctgtat ctggcggacc cgtggtggaa
      661 ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg tcccagggac tttgggggcc
      721 gtttttgtgg cccgacctga ggaagggagt cgatgtggaa tccgaccccg tcaggatatg
      781 tggttctggt aggagacgag aacctaaaac agttcccgcc tccgtctgaa tttttgcttt
      841 cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg ctgcagcatc gttctgtgtt
      901 gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta gggccagact gttaccactc
      961 ccttaagttt gaccttaggt cactggaaag atgtcgagcg gatcgctcac aaccagtcgg
     1021 tagatgtcaa gaagagacgt tgggttacct tctgctctgc agaatggcca acctttaacg
     1081 tcggatggcc gcgagacggc acctttaacc gagacctcat cacccaggtt aagatcaagg
     1141 tcttttcacc tggcccgcat ggacacccag accaggtccc ctacatcgtg acctgggaag
     1201 ccttggcttt tgacccccct ccctgggtca agccctttgt acaccctaag cctccgcctc
     1261 ctcttcctcc atccgccccg tctctccccc ttgaacctcc tcgttcgacc ccgcctcgat
     1321 cctcccttta tccagccctc actccttctc taggcgccgg ccggatccca gtgtggtggt
     1381 acgtaggaat tcgccaccat gggcagacac ctcctgctgc tcctgttcag catcctgtac
     1441 atgctgctgg cccaggccaa ggacaacacc gtgcccctga agctgatcgc cctgctggcc
     1501 aacggcgagt tccacagcgg cgagcagctg ggcgagaccc tgggcatgag cagagccgct
     1561 atcaacaagc acatccagac cctgagagac tggggcgtgg acgtgttcac cgtgcccggc
     1621 aagggctaca gcctgcccga gcccatccag ctgctgaacg ccaagcagat cctgggccag
     1681 ctggacggcg gaagcgtggc cgtgctgccc gtgatcgaca gcaccaacca gtacctgctg
     1741 gacagaatcg gcgagctgaa gagcggcgac gcctgcatcg ccgagtacca gcaggccggc
     1801 agaggcagaa gaggcagaaa gtggttcagc ccctttggcg ccaacctgta cctgagcatg
     1861 ttctggagac tggagcaggg ccccgcagcc gcaatcggcc tgagcctggt gatcggcatc
     1921 gtgatggccg aggtgctgag aaagctgggc gccgacaagg tgagagtgaa gtggcccaac
     1981 gacctgtacc tgcaggacag aaagctggcc ggcatcctgg tggagctgac cggcaagacc
     2041 ggcgacgccg cacagatcgt gatcggcgcc ggcatcaaca tggccatgag aagagtggag
     2101 gaaagcgtgg tgaaccaggg ctggatcacc ctgcaggagg ccggcatcaa cctggacaga
     2161 aacaccctgg cagccatgct gatcagagag ctgagagccg cactggagct gttcgagcag
     2221 gagggcctgg ctccctacct gagcagatgg gagaagctgg acaacttcat caacagaccc
     2281 gtgaagctga tcatcggcga caaggagatc ttcggcatca gcagaggcat cgacaagcag
     2341 ggcgccctgc tgctggagca ggacggcatc atcaagccct ggatgggagg cgagatcagc
     2401 ctgagaagcg ccgagaaggg ccccgactac aaggacgatg atgacaagtg aggatccccc
     2461 gacctcgacc ctggctaaag aattcgccag cacagtggtc gaccctgtgg aatgtgtgtc
     2521 agttagggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2581 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     2641 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2701 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2761 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2821 ttggaggcct aggcttttgc aaaaagctta ccatgaccga gtacaagccc acggtgcgcc
     2881 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     2941 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     3001 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     3061 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     3121 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     3181 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     3241 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     3301 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     3361 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     3421 cctggtgcat gacccgcaag cccggtgcct gacgcccgcc ccacgacccg cagcgcccga
     3481 ccgaaaggag cgcacgaccc catgcatcga taaaataaaa gattttattt agtctccaga
     3541 aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa gtaacgccat
     3601 tttgcaaggc atggaaaaat acataactga gaatagagaa gttcagatca aggtcaggaa
     3661 cagatggaac agctgaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
     3721 gctcagggcc aagaacagat ggaacagctg aatatgggcc aaacaggata tctgtggtaa
     3781 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tccagccctc
     3841 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     3901 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     3961 cccgagctca ataaaagagc ccacaacccc tcactcgggg cgccagtcct ccgattgact
     4021 gagtcgcccg ggtacccgtg tatccaataa accctcttgc agttgcatcc gacttgtggt
     4081 ctcgctgttc cttgggaggg tctcctctga gtgattgact acccgtcagc gggggtcttt
     4141 cacatgcagc atgtatcaaa attaatttgg ttttttttct taagtattta cattaaatgg
     4201 ccatagttgc attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc
     4261 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4321 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4381 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4441 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4501 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4561 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4621 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4681 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4741 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4801 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4861 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     4921 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     4981 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5041 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5101 gtcatgagat tatcaaaaag gatcttcacc tagatccttt tgcggccgca aatcaatcta
     5161 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     5221 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     5281 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     5341 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     5401 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     5461 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     5521 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     5581 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     5641 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     5701 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     5761 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     5821 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     5881 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     5941 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     6001 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     6061 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     6121 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccac
//
